Exploring the role of HINT1 protein in neuronal function

描述（由申请人提供）：蛋白激酶C相互作用蛋白（PKCI/HINT1）是一种小蛋白，属于组氨酸三联体（HIT）蛋白家族。死后研究已经确定PKCI/HINT1是精神分裂症和双相情感障碍的候选分子。在我们实验室的研究中，PKCI/HINT1 KO小鼠表现出一种行为和内分泌特征表型，表明情绪功能的变化，包括焦虑样和抗抑郁样行为以及对安非他明的超敏感性。这些结果提示PKCI/HINT1在正常中枢神经系统功能和精神疾病状态中起关键作用。目前的R21提案旨在为在细胞水平上表征PKCI/HINT1功能的长期研究奠定基础，并开发新的蛋白质抑制剂以促进表征，并有可能开发成相关中枢神经系统疾病的新型治疗剂。具体目标包括：目标1。发展PKCI/HINT1的细胞检测，关注与神经元存活、神经元生长和钙信号相关的潜在活动。分析发展将强调高通量能力，以促进未来的药物发现和优化工作。目标2。使用基于结构的计算药物设计方法鉴定具有高概率结合PKCI/HINT1的低分子量化合物。然后使用体外荧光和ITC分析，并辅以核磁共振分析，分析鉴定的化合物结合PKCI/HINT1的能力。从本质上讲，所提出的工作将填补PKCI/HINT1的已知三维结构与其已知的体内行为特征之间的空白。在本研究中获得的关于PKCI/HINT1细胞功能的知识将填补这一空白，允许对PKCI/HINT1的药理学进行广泛的未来研究，可能导致合理的药物设计方法，以开发新的治疗情绪障碍的药物。

DESCRIPTION (provided by applicant): Protein kinase C interacting protein (PKCI/HINT1) is a small protein belonging to the histidine triad (HIT) family of proteins. Postmortem studies have identified PKCI/HINT1 as a candidate molecule for schizophrenia and bipolar disorder. In studies from our laboratory, PKCI/HINT1 KO mice displayed a phenotype of behavioral and endocrine features which indicate changes of mood function, including anxiolytic- like and anti-depressant like behaviors and supersensitivity to amphetamine. These results suggest that PKCI/HINT1 plays a key role in normal CNS function and state of mental diseases. The present R21 proposal is designed to set the groundwork for long-term studies characterizing the function of PKCI/HINT1 at the cellular level and developing novel inhibitors of the protein to facilitate the characterization as well as have the potential to be developed into novel therapeutic agents for related CNS disorders. The specific aims include: Aim 1. Develop cellular assays for PKCI/HINT1 focusing on potential activities associated with neuronal survival, neuronal growth and calcium signaling. Assay development will emphasize high- throughput capabilities to facilitate future drug discovery and optimization efforts. Aim 2. Identify low molecular weight compounds with a high probability of binding to PKCI/HINT1 using structure-based computational drug design methods. Identified compounds will then be assayed for their ability to bind PKCI/HINT1 using in vitro fluorescence and ITC assays, supplemented with NMR analysis. In essence, the proposed work will fill a gap between the known 3D structure of PKCI/HINT1 and its known in vivo behavioral characteristics. Knowledge of the cellular function of PKCI/HINT1 gained in the proposed study will fill this gap allowing for extensive future studies of the pharmacology of PKCI/HINT1, potentially leading to a rational drug design approach to the development of novel therapeutic agents for mood disorders.